HuC Double Nickase Plasmid (h): sc-400172-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HuC Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- HuC Double Nickase Plasmid (h) and HuC Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ELAVL3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HuC Antibody (D-6): sc-515624
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HuC Double Nickase Plasmid (h) | sc-400172-NIC | 20 µg | $410.00 | |||
HuC Double Nickase Plasmid (h2) | sc-400172-NIC-2 | 20 µg | $410.00 |
ELAVL3 (HuC) is a neuron-enriched RNA-binding protein that recognizes AU-rich elements in target transcripts and regulates post-transcriptional gene expression through control of mRNA stability, localization, and translation. It contributes to neuronal differentiation and maintenance by shaping RNA regulons linked to synaptic function and cytoskeletal dynamics, and it interfaces with broader RNA processing pathways including alternative splicing and stress-responsive RNA granule biology. Altered neuronal RNA-binding protein activity and downstream transcriptomic programs are frequently studied in the context of neurodevelopmental and neurodegenerative mechanisms, where ELAVL3 serves as a marker and functional node in neuron-specific gene regulation. Human ELAVL3 research commonly examines how HuC-dependent RNA control influences neuronal maturation, excitability, and vulnerability under cellular stress.
HuC Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ELAVL3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ELAVL3. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt ELAVL3 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of ELAVL3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.